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* Corresponding author 1 SPRAM - UMR 5819 - Structures et propriétés d-architectures moléculaire 2 INSERM U823, équipe 8 Immunologie Analytique des Pathologies Chroniques Institut d-oncologie-développement Albert Bonniot de Grenoble

Abstract : The secretions of molecules by cells are of tremendous interest for both fundamental insights studies and medical purposes. In this study, we propose a new biochip-based approach for the instantaneous monitoring of protein secretions, using antibody production by B lymphocytes cultured in vitro. This was possible thanks to the Surface Plasmon Resonance imaging SPRi of a protein biochip where antigen proteins Hen Egg Lysozyme, HEL were micro-arrayed along with series of control proteins. B cell hybridomas were cultured on the chip and the secretion of immunoglobulins antibody specific to HEL was monitored in real-time and detected within only few minutes rather than after a 30-60 min incubation with standard ELISA experiments. This fast and sensitive detection was possible thanks to the sedimentation of the cells on the biochip sensitive surface, where local antibody concentrations are much higher before dilution in the bulk medium. An other interesting feature of this approach for the secretion monitoring was the independence of the SPR response-after normalization-regarding to the density of the surface-immobilized probes. Such biosensor might thus pave the way to new tools capable of both qualitative and semi-quantitative analysis of proteins secreted by other immune cells.

Keywords : Protein micro-array Surface plasmon resonance imaging Real-time detection Cellular secretion Blood cell

Autor: Sarah Milgram - Sandra Cortes - Marie-Bernadette Villiers - Patrice Marche - Arnaud Buhot - Thierry Livache - Yoann Roupioz -

Fuente: https://hal.archives-ouvertes.fr/


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